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P Challa, P Gonzalez, RR Allingham, DL Epstein, C Bowes Rickman; Identification of Genes Differentially Expressed in Individuals With Pseudoexfoliation Syndrome . Invest. Ophthalmol. Vis. Sci. 2002;43(13):4025.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To identify genes differentially expressed in the lens capsules of normal and pseudoexfoliation (PEX) specimens. Methods: Duke University IRB approval and patient consent was obtained. Surgical lens capsules (lens epithelia) from PEX individuals and normal age matched controls undergoing routine cataract extraction were collected and immediately stored on dry ice. Total RNA was extracted in TRIzol plus glycogen and DNase digested. Because of the small size of the samples, representative full-length, high fidelity cDNA pools were synthesized by the SMART PCR cDNA method. Human Life Grid 1.0 unigene macroarrays were differentially screened with lens cDNA probes derived from these SMART cDNA pools. Differential expression of genes identified on the macroarrays was confirmed by quantitative real-time PCR. Results: Eight of the cDNA clones represented in the array hybridized more intensely to the PEX-derived probe than to the normal controls. These PEX candidate genes represent currently uncharacterized genes. Quantitative analyses, by real-time PCR, of the transcript levels confirm that expression of these DNAs is elevated in the lens epithelium of the PEX derived samples. Conclusion: In vivo gene expression data can be successfully obtained on surgically excised lens capsule specimens despite their small size. We have identified distinct differences between gene expression in PEX and normal lens tissue. Understanding differential gene expression between PEX and normal lens capsules is important to unraveling the pathogenesis of pseudoexfoliation.
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